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Technical Abstract: Assessing the total dietary intake of vitamins from all sources is a current topic of interest in the field of human nutrition. Such an assessment would be aided considerably by having rapid and accurate analytical methodology that can be applied to vitamin analysis of foods and dietary supplements. It is desirable also to have methods which are capable of determining several vitamins simultaneously. Important sources of vitamin intake today include dietary supplements, such as multivitamin tablets, and vitamin-fortified foods. In these sources, vitamins typically are added in a single chemical form, and matrix issues are generally not as complex as those found when determining the natural vitamin content of foods. We have developed LC-UV and LC-MS methods for the simultaneous determination of several water-soluble vitamins in multivitamin and multivitamin/multi-mineral tablets and are now extending these methods to analysis of fortified breakfast cereals. We will present data for thiamine, niacin, pyridoxine, pantothenic acid, folic acid, biotin, riboflavin, and ascorbic acid, including assessment of accuracy and precision. These data will be used by public and private sector analytical laboratories engaged in vitamin determinations.